This proposal describes a program for developing multidimensional analytical techniques for the analysis of complex biological mixtures that are extracted from Drosophila melanogaster as a function of the age of the organism. Preliminary data show that a new gas-phase ion mobility/time-of-flight mass spectrometry technique can be coupled to other separation techniques in order to separate and characterize complex mixtures of tryptic peptides obtained from digestion of soluble proteins. The gas-phase separation reduces spectral congestion and chemical noise associated with mass spectrometry alone, provides additional information about charge state and amino acid composition, and allows a parallel approach for obtaining fragmentation data (an important factor for assigning peptide sequence and identifying proteins). This proposal describes the further advancement of these technologies and their applications to the emerging field of proteomics. An aggressive plan that aims to map the proteome for the fundamental model Drosophila organism is proposed. This existence of a detailed proteomic map for this system is an important scientific step that will allow a range of genetic models to be investigated in detail. These models are chosen to display characteristics of diseased states (e.g., neurodegenerative conditions) for which genetic studies in humans are intractable.